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Volume 2, Issue2, April 2011
International Journal Of Pharma Professional’s
Research
Review Article
HPLC: A VERSATILE CHROMATOGRAPHIC
APPROACH USED FOR QUALITATIVE AND
QUANTITATIVE PURPOSES- A REVIEW
VIKRANT ARYA1*
, ANKUR BHARDWAJ2, VINIT SHARMA
3 ISSN NO:0976-6723
Department of Pharmacognosy, Amar Shaheed Baba Ajit Singh Jujhar Singh Memorial Post Graduate College of
Pharmacy, Bela, Ropar, Punjab-140111, India
Abstract High Performance Liquid Chromatography (HPLC) has been used for the analysis of natural and synthetic
compounds. This review gives an overview about some special features involving the mode of operation, mobile
phase, detector, column and the flow rate of solvent system employed. This review paper provides useful
information regarding the identification, separation and quantification of various natural as well as synthetic
compounds.
Keywords: - : HPLC, quantification, phytoconstituents, synthetic
Introduction A variety of methods are available for analyzing
pharmaceutical compounds. High Performance/
Pressure Liquid Chromatography (HPLC) is one of
the best methods of choice for analysing a variety
of natural and synthetic compounds. It is because it
offers high performance over ambient pressure.[1]
This method is used for checking the purity of new
drug candidates, monitoring changes or in process
testing for developing new formulations and quality
control or assurance of final drug products. HPLC
is a dynamic adsorption process[2] and is a
separation technique conducted in the liquid phase
in which a sample is separated into its constituent
components by distributing between the mobile
phase and a stationary phase. HPLC utilises a liquid
mobile phase to separate the components of a
mixture. The stationary phase can be a liquid or a
solid phase. These components are first dissolved in
a solvent, and then forced to flow through a
chromatographic column under a high pressure.[3] Correspondence Address: Vikrant arya
Amar Shaheed Baba Ajit Singh Jujhar Singh
Memorial Post Graduate College of Pharmacy,
Bela, Ropar, Punjab-140111, India
Email: [email protected]
Phone:91-09736105832
HPLC acquires a high degree of versatility not found in other
chromatographic systems and it has the ability to easily
separate a wide variety of chemical mixtures. Some axioms
of HPLC are: Sample must be soluble; for separation,
analytes must be retained and have differential migration in
the column; the mobile phase controls HPLC separation; the
final analyte solution should be prepared in the mobile
phase.[4]
In this review, we discussed the role of HPLC in the
qualitative and quantitative determination of natural
phytoconstituents and synthetic drug molecules.
HPLC EVALUATION FOR PHYTOCONSTITUENTS
[5-45]
For quality control of herbal products, high performance
liquid chromatography (HPLC) is a popular method for the
analysis of herbal medicines because it is accurate, precise
and not limited by the volatility or stability of the sample
compounds. HPLC combined with diode array detector
(HPLC/DAD), electrochemical detection (HPLC-ED), mass
spectrometer (HPLC /MS) have been successfully employed
in qualitative and quantitative determination of various types
phytoconstituents including alkaloids, flavonoids, tannins,
glycosides, triterpenes, sterols etc. A brief description of
HPLC conditions for some phytoconstituents were discussed
in table 1.
Available Online at www.ijppronline.in
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Table 1: HPLC conditions for some
phytoconstituents evaluation[5-45]
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PHYTOCONSTITUENTS EVALUATED VIA HPLC[5-45]
Large number of phytoconstituents were quantified via
HPLC technique for example: flavonoids (quercetin,
kaempferol, cyanidin); flavonoid glycosides (rutin); tannins
(ellagic acid, ellagitannins); alkaloids (camptothecin,
phyllanthin); polyphenols (arbutin); coumarins; triterpenes
(ursolic acid, arjunolic acid); carotenoids (lycopene);
sphingolipids etc. Some of reported quantified
phytoconstituents were shown in figure 1.
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Figure 1: Phytoconstituents quantified via HPLC
HPLC EVALUATION FOR SYNTHETIC
DRUGS [46-86]
HPLC methods have been used in the determination
of drugs in biological fluids and in pharmaceutical
dosage forms. HPLC determination method with
spectroscopic detection is useful for routine quality
control of drugs in pharmaceutical formulations and
stability studies. Various types of detectors: photo
diode array, ultraviolet (UV), ultraviolet visible
(UV-VIS), fluorometric, fluorescence have been
employed along with different types of columns (C-
8, RP-8, RP C-18, SB-CN, ODS-2) depending upon
the synthetic drug to be evaluated. Some of reported
synthetic drug molecules evaluated via HPLC were
shown in table 2 given below.
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SYNTHETIC MOLECULES EVALUATED
VIA HPLC [46-86]
Synthetic molecules were successfully evaluated
both qualitatively as well as quantitatively via
HPLC as shown in figure 2. Several
pharmaceutically important Antiviral drugs:
lamivudine, stavudine, nevirapine, zidovudine;
NSAIDS: acetaminophen, meloxicam, paracetamol,
phenylbutazone; Antibiotics: amoxicillin, cefepime,
rifampicin; CNS stimulants: amphetamine; Beta
blockers: atenolol; Preservatives: benzalkonium
chloride; Diuretics: hydrochlorothiazide;
Antidiabetics: metformin; Anticancer:
methotrexate; Corticosteroids: prednisone,
betamethasone, dexamethasone; Antianginal:
Lercanidipine, Nicorandil; Immunomodulator :
azathioprine; Anti-allergic: cetirizine;
Antidepressant: Nitroxazepine, milnacipran;
Veterinary anesthetic: Ketamine; Anti-
inflammatory: sodium ozagrel; Secretolytic:
Ambroxol were successfully evaluated via HPLC.
Figure 2: Synthetic drug molecules evaluated via HPLC
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CONCLUSION
HPLC is one mode of chromatography, one of the
most used analytical techniques. HPLC applications
can be used effectively for screening analysis as
well as quality evaluation of natural as well as
synthetic compounds. Owing to the simplicity and
efficiency of HPLC specific and rapid
determination of various natural and synthetic
compounds can be carried out. HPLC can be
employed for the routine analysis of natural and
synthetic compounds in pharmaceutical
formulations and in bulk drug preparations as well
as for the quality assurance of related extracts and
market samples. Interest in HPLC has increased
with improvements in its instrumentation and
methods and especially in the last few years, with
the combination of hyphenated techniques like
HPLC–MS/MS, HPLC/DAD/MS, HPLC-ED.
REFERENCES
1).Ahuja S and Dong MW. Handbook of
Pharmaceutical Analysis by HPLC. 1st ed.
Academic Press Publisher.UK 2005.
2).Ahuja S and Alsante K. Handbook of Isolation
and Characterization of Impurities in
Pharmaceuticals. Academic Press New York 2003.
3).Snyder LR and Kirkland JJ. Introduction to
Modern Liquid Chromatography. John Wiley and
Sons New York 1979.
4).Meyer VR. Practical HPLC. Wiley New York
1998.
5).Correia H et al. Polyphenolic profile
characterization of Agrimonia eupatoria L. by
HPLC with different detection devices. Biomedical
Chromatography 2006; 20: 88-94.
6).Aaby K et al. Characterization of Phenolic
Compounds in Strawberry (Fragaria × ananassa)
Fruits by Different HPLC Detectors and
Contribution of Individual compounds to Total
Antioxidant Capacity. Journal of Agricultural and
Food Chemistry 2007; 55: 4395-4406.
7).Pothitirat W and Gritsanapan W. HPLC
Quantitative Analysis Method for the
Determination of α-Mangostin in Mangosteen Fruit
Rind Extract. Thai Journal of Agricultural Science
2009; 42(1): 7-12.
8).Mousavinejad G et al. Identification and
quantification of phenolic compounds and their
effects on antioxidant activity in pomegranate juices of eight
Iranian cultivars. Food Chemistry 2009; 115: 1274–1278.
9).Kittipongpatana N et al. High-Performance Liquid
Chromatographic Method for Separation and Quantitative
Analysis of Arbutin in Plant Tissue Cultures. CMU. Journal
of Natural Science 2007; 6 (1): 65.
10).Rajalakshmi PV and Senthil KK. Direct HPLC analysis
of quercetin in exudates of Abutilon indicum (Linn).
Malvaceae. Journal of Pharmaceutical Science and
Technology 2009: 1 (2): 80-83.
11).Stanic G et al. HPLC Analysis of Esculin and Fraxin in
Horse-Chestnut Bark (Aesculus hippocastanum L.). Croatica
Chemica Acta 1999; 72 (4): 827-834.
12).Qiao C et al. HPLC determination of two bioactive
flavone glycosides and GC-MS analysis of volatile oil
constituents in Alpinia nigra. Asian Journal of Traditional
Medicines 2007; 2 (3): 85-91.
13).Gazdik Z et al. Use of Liquid Chromatography with
Electrochemical Detection for the Determination of
Antioxidants in Less Common Fruits. Molecules 2008; 13:
2823-2836.
14).Samee W and Vorarat S. Simultaneous Determination of
Gallic acid, Catechin, Rutin, Ellagic Acid and Quercetin in
Flower Extracts of Michelia alba, Caesalpinia pulcherrima
and Nelumbo nucifera by HPLC. Journal of Thai Pharm
Health Science; 2007: 2(2):131-137.
15).Liu X et al. Quantification and Purification of Mulberry
Anthocyanins with Macroporous Resins. Journal of
Biomedicine and Biotechnology 2004; 5 (2004) 326–331.
16).Montoro P et al. Identification and Quantification of
Components in Extracts of Uncaria tomentosa by HPLC-
ES/MS. Phytochemical analysis 2004; 15: 55-64.
17).Singh DV et al. RP-LC determination of oleane
derivatives in Terminalia arjuna. Journal of Pharmaceutical
and Biomedical Analysis 2002; 28: 447–452.
18).Lee J et al. Antibacterial effects of S-(-)-tulipalin B
isolated from Spiraea thunbergii Sieb. on Escherichia coli, a
major food borne pathogenic microorganism. Journal of
Medicinal Plants Research 2008; 2(3): 059-065.
19).Wang L et al. HPLC Quantification of Sphingolipids in
Soybeans with Modified Palmitate Content. Journal of
Agriculture and Food Chemistry 2006.
20)Tsujikawa K et al. Determination of salvinorin A and
salvinorin B in Salvia divinorum-related products circulated
in Japan. Forensic Science International 2008; 180: 105–109.
21).Mertz C et al. Analysis of Phenolic Compounds in Two
Blackberry Species (Rubus glaucus and Rubus adenotrichus) by High-Performance Liquid Chromatography with Diode Array
Detection and Electrospray Ion Trap Mass Spectrometry.
252
Volume 2, Issue2, April 2011
Journal of Agriculture and Food Chemistry 2007;
55: 8616–8624.
22).Qiao CF. HPLC Analysis of Bioactive
Diterpenoids from the Root Bark of Pseudolarix
kaempferi. Journal of Food and Drug Analysis
2006; 14 (4): 353-356.
23).Olszewska M. Quantitative HPLC analysis of
flavonoids and chlorogenic acid in the leaves and
inflorescences of Prunus serotina. Acta
chromatographica 2007; 19: 253-269.
24).Burkhardt S et al. Detection and Quantification
of the Antioxidant Melatonin in Montmorency and
Balaton Tart Cherries (Prunus cerasus). Journal of
Agriculture and Food Chemistry 2001; 49: 4898-
4902.
25).Tripathi AK et al. Quantitative Determination
of Phyllanthin and Hypophyllanthin in Phyllanthus
Species by High-performance Thin Layer
Chromatography. Phytochemical Analysis 2006;
17: 394-397.
26).Frye A and Haustein C. Extraction,
identification, and quantification of Harmala
Alkaloids in Three Species of Passiflora. American
Journal of Undergraduate Research 2007; 6 (3): 19-
26.
27).Suchomelova J et al. HPLC quantification of
seven quaternary benzo[c]phenanthridine alkaloids
in six species of the family Papaveraceae. Journal
of Pharmaceutical and Biomedical Analysis 2007;
44: 283–287.
28).Castellanos-Santiago E and Yahia EM.
Identification and Quantification of Betalains from
the Fruits of 10 Mexican Prickly Pear Cultivars by
High-Performance Liquid Chromatography and
Electrospray Ionization Mass Spectrometry. Journal
of Agriculture and Food Chemistry 2008; 56: 5758-
5764.
29).Ergonul PG and Nergiz C. Determination of
Organic Acids in Olive Fruit by HPLC. Czech
Journal of Food and Science 2010; 28 (3): 202-205.
30).Silva MGV et al. Variation of Ursolic Acid
Content in Eight Ocimum Species from
Northeastern Brazil. Molecules 2008; 13: 2482-
2487.
31).Wu SF. et al. Camptothecinoids from the seeds
of Taiwanese Nothapodytes foetida. Molecules
2008; 13: 1361-1371.
44).Rangkadilok N et al. Identification and
Quantification of Polyphenolic Compounds in
32).Gbaguidi F et al. HPLC quantification of two isomeric
triterpenic acids isolated from Mitracarpus scaber and
antimicrobial activity on Dermatophilus congolensis. Journal
of Pharmaceutical and Biomedical Analysis 2005; 39: 990–
995.
33).Perez OP et al. Isolation and characterisation of active
compounds from Mammea americana Lin. Revista cubanade
quimica 2007; 19 (1): 74-77.
34).Bicanic D et al. Quantification of lycopene in tomato
products: comparing the performances of a newly proposed
direct photothermal method and high-performance liquid
chromatography. Journal of the Science of Food and
Agriculture 2005; 85: 1149-1153.
35).Adhyapak S. High Performance Liquid Chromatographic
method for quantization of apigenin from dried root powder
of Gmelina arborea Linn. International Journal of Pharma
and Bio Sciences 2011; 2(1): 742-749.
36).Syamsudin et al. HPLC Analysis and pharmacokinetic
study of Mangostin after orally administration in rats. The
Pharma Research 2009; 2: 43-49.
37).Moriyama H et al. HPLC quantification of kaempferol-
3-Ogentiobioside in Cassia alata. Fitoterapia 2003; 74: 425–
430.
38).Qiao C et al. HPLC determination of two bioactive
flavone glycosides and GC-MS analysis of volatile oil
constituents in Alpinia nigra. Asian Journal of Traditional
Medicines 2007; 2 (3): 85-91.
39).Samee W and Vorarat S. Simultaneous Determination of
Gallic acid, Catechin, Rutin, Ellagic Acid and Quercetin in
Flower Extracts of Michelia alba, Caesalpinia pulcherrima
and Nelumbo nucifera by HPLC. Journal of Thai Pharm
Health Science 2007; 2(2): 131-137.
40).Fan JP and He CH. Simultaneous quantification of three
major bioactive triterpene acids in the leaves of Diospyros
kaki by high-performance liquid chromatography method.
Journal of Pharmaceutical and Biomedical Analysis 2006;
41: 950–956.
41).Chandrika UG et al. Identification and hplc
quantification of carotenoids of the fruit pulp of
Chrysophyllum roxburghii. Journal of National Science
Foundation of Sri Lanka 20005; 33 (2): 93-98.
42).Wallis IR et al. Quantification of sideroxylonals in
Eucalyptus Foliage by High performance Liquid
Chromatograpy. Phytochemical Analysis 2003; 14: 360-365.
43).Mulabagal V. et al. Cultivars of Apple Fruits That Are
Not Marketed with Potential for Anthocyanin Production.
Journal of Agriculture and Food Chemistry 2007; 55: 8165-
8169.
Longan (Euphoria longana Lam.) Fruit. Faculty of Science,
253
Volume 2, Issue2, April 2011
Mahidol University 2004.
45).Amna T et al. Determination and quantification
of camptothecin in an endophytic fungus by liquid
chromatography – positive mode electrospray
ionization tandem mass spectrometry. Current
science 2006; 91 (2): 208-212.
46).Schuman M et al. HPLC analysis of generic
antiretroviral drugs purchased in Rwanda. Bull.
Soc. Sci. Med. 2005; 3: 317-324.
47).Cole MD et al. 4-Bromo-2, 5-
dimethoxyphenethylamine (2C-B): A review of the
public domain literature. Science and Justice 2002;
42(4): 223-224.
48).Suzen S et al. Quantification of Acetaminophen
in Pharmaceutical Formulations Using High
Performance Liquid Chromatography. J. Fac.
Pharm. Ankara 1998; 27(2): 93-100.
49).Pires LR et al. HPLC determination of
amoxicillin comparative Bioavailability in healthy
volunteers after a single dose administration.
Journal of Pharmaceutical Science 2003; 6(2): 223-
230.
50).Pavlova V and Jovanovic SP. Simultaneous
Determination of Amphetamine, Methamphetamine
and Caffeine in Seized Tablets by High
Performance Liquid Chromatography. Acta
Chromatographia 2007; 18: 157-167.
51).Zaveri M and Khandhar A. Development and
Validation of a RP-HPLC for the Simultaneous
Estimation of Atenolol and Hydrochlorothiazide in
Pharmaceutical Dosage Forms. JPRHC; 2(3): 248-
252.
52).Mehta J et al. Development and Validation of a
Precise Method for Determination of Benzalkonium
Chloride (BKC) Preservative in Pharmaceutical
Formulation of Latanoprost Eye Drops. E-Journal
of Chemistry 2010; 7(1): 11-20.
53).Hurum et al. Determination of Cefepime and
Related Compounds Using HPLC with UV
Detection. LC GC Asia Pacific 2009; 121(2).
54).Sultana N et al. Simultaneous Determination of
Ceftriaxene Sodium and Non Steroidal Anti-
inflammatory Drugs in Pharmaceutical.
Formulations and Human Serum by RP-HPLC.
Journal of the Chinese Chemical Society 2010; 57:
1278-1285
55).Woodwad C and Major R. Preparative Purification of Corticosteroids by HPLC; Scalability
and Load ability using Agilent Prep C18 HPLC
Columns. Agilent Technologies, Publication USA 2005.
56).Popovic I et al. LC Determination of Lercanidipine and
its Impurities Using Drylab Software and Experimental
Design Procedures. Chromatographia 2008; 67: 449-454.
57).Basavaiah K et al. Quantitative determination of
Olanzapine in Pharmaceutical Preparations by HPLC. J.Mex.
Chem. Soc. 2008; 52(2): 120-124.
58).Kumar AKH et al. A Validated High Performance liquid
Chromatography Method for the Determination of rifampicin
and desacetyl rifampicin in plasma and urine. Indian Journal
Pharmacology 2004; 36(4): 231-233.
59).Jedziniak P et al. Determination of Phenylbutazone and
Oxyphenbutazone in Bovine Plasma Using High
Performance Liquid Chromatography with UV Detection.
Bull Vet Inst Pulawy 2005; 49: 223-226.
60).Altun ML. HPLC Method for the Analysis of
Paracetamol, Caffeine and Dipyrone. Turk J Chem 2002; 26:
521-528.
61).Bedor DCG et al. Development and Validation of a new
method for the quantification of Norflpxacin by HPLC-UV
and its application to a comparative pharmacokinetic study
in human volunteers. Brazilian Journal of Pharmaceutical
Sciences 2007; 43(2): 231-237.
62).Parveen PS et al. RP-HPLC Method for the Estimation
of Nitroxazepine Hydrochloride in Pharmaceutical Dosage
Forms. International Journal of Applied Biology and
Pharmaceutical Technology 2010; 1(3): 1181-1187.
63).Mehta PJ and Khatri DM. Development and Validation
of RP-HPLC Method for Determination of Milnacipran
Hydrochloride in Pharmaceutical Formulations. International
Journal of Pharmacy and Pharmaceutical Sciences 2010; 2:
137-141.
64).Yousefi G et al. Quantification of Polyethylene Glycol
esters of Methotrexate and Determination of their Partition
Coefficients by Validated HPLC Methods. Iranian Journal of
Pharmaceutical Research 2009; 8(1): 27-31.
65).Mehmood KT et al. Specific and Simple HPLC assay of
Ecofriendly Meloxicam in Pharmaceutical Formulations.
Journal of Pharmaceutical Sciences and Research 2010; 2:
878-883.
66).Jalalizadeh H et al. A Rapid and Sensitive HPLC method
for Determination of 2-Hydroxyflutamide in Human Plasma.
International Journal of Pharmacology 2006; 2(2): 221-225.
67).Sangiorgi E and Curatolo M. Rapid Determination of Six
Synthetic Corticosteroids in Urine by High Performance
Liquid Chromatograph Atmospheric Pressure Chemical
Ionisation Mass Spectometry. Istituto zooprofilattico
sperimentale delle venezie: 969-975.
68).Kwak HW and Amico DJD. Determination of
254
Volume 2, Issue2, April 2011
Dexamethasone sodium phosphate In the vitreous
by High Performance Liquid Chromatography.
Korcan J. Opthalmol 1995; 9: 79-83.
69).Yadav AH et al. RP-HPLC method for
Determination of Aceclofenac, Chloroxapine and
Paracetamol in Bulk and Pharmaceutical
Formulation. International Journal of
Pharmaceutical Research and Development 2009;
1(10): 1-7.
70).Reddy PV et al. RP-HPLC Determination of
Raloxifene in Pharmaceutical Tablets. Journal of
Chemistry 2006; 3(10): 60-64.
71).Bhaskar M et al. RP-HPLC method
Development for the Determination of Azathioprine
in Bulk drug and Pharmaceutical Dosage Forms.
International Journal of Chem.Tech Research 2010;
2(2): 1176-1179.
72).Yilmaz B. Reverse Phase HPLC Method for
Determination of Nebivolol in Pharmaceutical
Preparations. International Journal of
Pharmaceutical Science Review and Research
2010; 1(2): 14-17.
73).Yang JF et al. Determination of Gliclazide in
Human Plasma by High Performance Liquid
Chromatography. Asian Journal of Drug
Metabolism and Pharmacokinetics 2004; 4(3): 231-
234.
74).Arayne MS et al. Determination and
Quantification of Cetrizine HCl in Dosage
Formulations by RP-HPLC. Pakistan Journal of
Pharmaceutical Sciences 2005; 18(3): 7-11.
75).Barrett B et al. Validated HPLC-MS/MS
Method for Determination of Quetiapine in human
Plasma. Journal of Pharmaceutical and Biomedical
Analysis 2007; 44: 498-505.
76).Chitlange SS et al. Stability Indicating RP-
HPLC Method for Simultaneous Estimation of
Valsartan and Amlodipine in Capsule Formulation.
Asian J. Research Chem. 2008; 1(1): 15-18.
77).Reddy PB et al. Determination of Pantroprazole
sodium and Lansoprazole Individual Dosage Form
Tablets by RP-HPLC using Single Mobile Phase.
International Journal of Applied Biology and
Pharmaceutical Technology 2010; 1(2): 0976-4550.
78).Redmann S and Charles BG. A rapid HPLC Method
with fluorometric detection for determination of plasma
itraconazole and hydroxy-itraconazole concentration in
cystic fibrosis children with allergic bronchopulmonary
aspergillosis. Biomedical Chromatography 2005.
79).Shakya AK et al. Simple and Rapid HPLC Method for
the Determination of Alfuzosin in Human Plasma. Jourdan
Journal of Pharmaceutical Sciences 2010; 3(1): 25-35.
80).Lakkanatinaporn P and Matayatsuk C. Simultaneous
HPLC method for Determination of Sodium Trimethoprim
Phenylpropanol disulphonate and Sodium sulphaquinoxaline
in veterinary drugs. Songklanakarin J. Sci. Tecnol. 2004;
26(6): 849-854.
81).Kabra B et al. Simultaneous Quantitative Determination
of Zidovudine and nevirapine in human plasma using
isocratic, reverse phase high performance liquid
chromatography. Tropical Journal of Pharmaceutical
Research 2009; 8(1): 79-86.
82).Cheng ZZ et al. Identification and Determination of
Nicorandil and its Degradation Products by HPLC and
GC/MS. Chinese Chemical Letters 2006; 17(8): 1057-1060.
83).Pasha K et al. RP-HPLC method for determination of
Ambroxol in Pharmaceutical dosage forms. Research Journal
of Pharmaceutical, Biological and Chemical Sciences 2010;
1(3): 250.
84).Lakshmi KS et al. Simultaneous Determination of
Metformin and Pioglitazone by Reversed Phase HPLC in
Pharmaceutical Dosage Forms. International Journal of
Pharmacy and Pharmaceutical Sciences 2009; 1(2): 162-166.
85).Sun K et al. Determination of Sodium ozagrel in human
plasma and urine by HPLC with solid- phase extraction and
its application to pharmacokinetics. Asian Journal of
Pharmaceutical Sciences 2006; 1(3-4): 222-228.
86).Bolze S and Boulieu R. HPLC determination of
ketamine, norketamine and Dehydronorketamine in plasma
with a high purity reversed- phase sorbent. Clinical
Chemistry; 1998: 560-564.
255